Methods: In our experimental system, PBMC from healthy donors were stimulated for 4h with 50% sera from SLE patients. Total RNA was purified and expression of type I IFN-inducible genes, including DDX58, G1P2, MX1, OAS3, RSAD2, IFIT1, IFI35 were measured by real-time QRT-PCR analysis. Rage antibodies characterization: Anti-huRAGE reactivity was confirmed by ELISA and FACS using soluble RAGE and NSO stably cells transfected with full-length RAGE. Epitope mapping of anti-RAGE mAbs was performed using 293F cells expressing different human RAGE domain deletion mutants. Inhibition of the binding between RAGE and HMGB1/CpG complex was determined by ELISA.
Results: Anti-RAGE mAbs inhibited the IFNα gene signature by ~ 45 % which was comparable to the inhibition by soluble RAGE-Fc (54±11%). Interestingly, these mAbs all bind to RAGE at the C2 domain (plasma-membrane proximal Ig-loop domain). Furthermore, these mAbs also inhibit the binding between RAGE and HMGB1 complexed with CpG DNA.
Conclusion: These results suggest that RAGE and HMGB1 contribute to the activation of the mononuclear cells in SLE, and that antibodies targeting RAGE may present a novel therapeutic approach in the treatment of SLE.
B. Chen, Full, 3; S. Mao, MedImmune, Inc, 3; Y. Yao, 2250 shares, 1; MedImmune, Inc., 3; C. Chang, MedImmune, Inc, 3; G.P. Sims, MedImmune, Inc, 3; P. Chowdhury, yes, 1; Full-time, 3; L. Audoly, Yes, 1; Yes, 3; R. Herbst, yes, 1; yes, 3; H. Wu, yes, 1; yes, 3; B. Jallal, yes, 1; yes, 1; full, 3; P. Kiener, yes, MedImmune, 1; yes, 3; yes, Synovex, 6; A.J. Coyle, Yes, 1; Yes, 1; Full Time, 3; Yes, 6.
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